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Electrogenetic cellular insulin release for real-time glycemic control in type 1 diabetic mice

Data underlying the figures in the publication “Electrogenetic cellular insulin release for real-time glycemic control in type 1 diabetic mice”, published in Science, 2020, 368, 993-1001. <a href="https://science.sciencemag.org/content/368/6494/993">https://science.sciencemag.org/content/368/6494/993</a> Table of contents: 1. Electrogenetics source data file; File openable with Graphpad Prism containing the source data for the main Figures: 1-4 (Transgene expression by SEAP measurement), 5b, 6a, 6f (Insulin by ELISA), 5c, 6b, 6e (NanoLuc luciferase), 7c (NanoLuc luciferase in vivo) and 7a, 7d (Glycemia). Transgene expression by SEAP measurement Data for main Figures 1, 2, 3, 4. SEAP (human placental secreted alkaline phosphatase) levels were profiled in cell culture supernatants using a colorimetric assay. 100 µL 2x SEAP assay buffer (20 mM homoarginine, 1 mM MgCl2, 21% diethanolamine, pH 9.8) was mixed with 80 µL heat-inactivated (30 min at 65°C) cell culture supernatant. After the addition of 20 µL substrate solution (120 mM p-nitrophenyl phosphate; cat. no. AC128860100, Thermo Fisher Scientific), the absorbance time course was recorded for 45 min at 405 nm and 37°C using a Tecan Genios PRO plate reader (cat. no. P97084; Tecan Group AG, Maennedorf, Switzerland) and the SEAP levels were determined as follows: first, absorbance change over time (slope) was calculated. According to the Beer–Lambert’s law, absorbance is proportional to the concentration of a colored compound and depends on the light path length (d) and extinction coefficient (ε) (ε for p-nitrophenyl (εpNP) = 18.600 M−1 cm−1). Enzymatic activity EA [U/L] was calculated from the equation: EA = slope × dilution factor × εpNP−1 × d−1  Values in the file present determined SEAP levels. Insulin by ELISA Data for Figures 5b, 6a, 6f. Values in the file present Insulin level as determined by ELISA kit. The assay was performed according to manufacturer’s instructions. NanoLuc luciferase Data for Figures 5c, 6b, 6e. NanoLuc® luciferase was quantified in cell culture supernatants using the Nano-Glo® Luciferase Assay System (cat. no. N1110; Promega, Duebendorf, Switzerland). In brief, 7.5 µL of cell culture supernatant was added per well of a black 384-well plate and mixed with 7.5 µL substrate-containing assay buffer. Total luminescence was quantified using a Tecan Genios PRO plate reader (Tecan Group AG). Values in the file present measured luminescence levels. NanoLuc luciferase in vivo Data for Figure 7c. Aliquots of 15 µL of whole-blood samples were diluted in 5 μL of 50 mM EDTA and frozen at -20 °C until NanoLuc® quantification as described above. Values in the file present measured luminescence levels normalized to time point 0 (Normalization individually for each mouse). Glycemia Data for Figures 7a, 7d. Blood glucose level was determined using a glucometer (Contour®Next, Bayer Healthcare, Leverkusen, Germany). Values in the file present measured glucose levels. 2. Figure 7b; Data for Figure 7b. File openable with Graphpad Prism. Blood glucose level was determined using a glucometer (Contour®Next, Bayer Healthcare, Leverkusen, Germany). Values in the file present measured glucose levels. 3. Figure 6c and 6d; Excel file with the data for Figures 6c, 6d. NanoLuc® luciferase was quantified in cell culture supernatants using the Nano-Glo® Luciferase Assay System (cat. no. N1110; Promega, Duebendorf, Switzerland). In brief, 7.5 µL of cell culture supernatant was added per well of a black 384-well plate and mixed with 7.5 µL substrate-containing assay buffer. Total luminescence was quantified using a Tecan Genios PRO plate reader (Tecan Group AG). Values in the file present measured luminescence levels.

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32/100

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Stewardship: 8
Harmonization: 4
Access: 16
Reuse readiness: 4
Engagement: 0

Topics

Electrogenetics, Synthetic Biology, Bioengineering